Expression of Protein Antigen Hiv -1 P24 in Transgenic Tobacco Plants

Only available on StudyMode
  • Download(s) : 61
  • Published : June 7, 2011
Open Document
Text Preview
VIETNAM NATIONAL UNIVERSITY – HOCHIMINH CITY
INTERNATIONAL UNIVERSITY
School of Biotechnology

EXPRESSION OF PROTEIN ANTIGEN HIV -1 P24 IN TRANSGENIC TOBACCO PLANTS

A thesis submitted to
The School of Biotechnology, International University
In partial fulfillment of the requirements for the degree of B.S. in Biotechnology

Student name: Nguyen Ngoc Tu Anh – BT050036
Supervisor: Dr. Nguyen Thi Thanh
Dr. Phan Dinh Phap

June/2010
ABSTRACT
The production of antigens for vaccines in plants has the potential as a safe and cost-effective alternative to traditional production systems. Transgene expression from the plant’s plastid genome represents a promising strategy in molecular farming because of the plastid’s potential to accumulate foreign proteins to high levels and the increased biosafety provided by the maternal mode of organelle inheritance. In this thesis, we confirm the high-level expression of HIV-1 p24 antigen in transgenic tobacco plants through plastid transformation. PCR analysis confirmed the presence of the HIV-1 p24 sequence within the chloroplast genome of transgenic lines. SDS-PAGE gel electrophoresis of protein extracts from transgenic plants identified plant-expressed HIV-1 p24 protein. Quantification of the recombinant protein HIV-1 p24 using Aligent 2100 Bioanalyzer estimated yields of about 13 mg per g of soluble leaf protein. Our results indicate that plant-based transgenic expression represents the great potential of transgenic plastids to develop HIV vaccine components at low cost and high yield and for use in HIV diagnostic procedures.

ACKNOWLEDGMENT
First and foremost, I would like to thank to Dr. Nguyen Thi Thanh for the valuable guidance and advice to complete my thesis for Bachelor of Science degree. I also would like to thank to Dr. Phan Dinh Phap for his help and introduction to the Institute of Tropical Biology. Besides, I would like to thank the authority of the Institute of Tropical Biology for providing me with a good environment and facilities to complete this thesis. I also would like to thank to Dr. Nguyen Huu Ho, Mr. Le Tan Duc, Mr. Loc, Mr. Tri, Mr. Duong and especially Mr. Huan who helped me a lot in doing experiments and gave me advantageous conditions to complete my thesis. In addition, I would like to acknowledge the School of Biotechnology International University for giving me supports during the time I studied here. Finally, an honorable mention goes to our families and friends for their understandings and supports on us in completing this thesis. Without helps of the particular that mentioned above, I would face many difficulties while doing this thesis.

TABLE OF CONTENTS
ABSTRACTi
ACKNOWLEDGMENTii
LIST OF TABLESvi
LIST OF FIGURESvii
ABBREVIATIONSviii
CHAPTER 1.INTRODUCTION2
CHAPTER 2.LITERATURE REVIEW5
2.1TOBACCO PLANTS5
2.1.1Geographical distribution of tobacco plants5
2.1.2Botanical description of tobacco plants6
2.1.2.1Stems6
2.1.2.2Leaves6
2.1.2.3Flowers7
2.1.2.4Roots7
2.1.2.5Seeds7
2.1.3Roles of tobacco plants in the life8
2.2HUMAN IMMUNODEFICIENCY VIRUS TYPE 1(HIV-1)10
2.2.1Structure and genome of HIV-110
2.2.2Replication cycle of HIV-112
2.2.3The potential of the p24 capsid protein as a vaccine antigen13
2.2.4Facts of HIV in Viet Nam15
2.3PLASTID TRANSFORMATION16
2.3.1Plastid and their genome structure16
2.3.2Advantages and disadvantages of plastid transformation over nuclear transformation16
2.4MOLECULAR FARMING - PLANTS AS EXPRESSION SYSTEM18
2.5SDS-PAGE AND PROTEIN ASSAY20
2.5.1Protein assay theory20
2.5.2Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE)20
2.6BIOANALYZER AND PROTEIN 50 ASSAY22
2.6.1Agilent 2100 bioanalyzer22
2.6.2Protein 50 LabChip kit23
CHAPTER 3.MATERIAL AND METHOD26...
tracking img